The present invention relates to Published European Patent Application 108 565 relates to compounds of the General Formula: 
and their pharmaceutically acceptable salts, in which R1 is an aliphatic hydrocarbon radical having 8-30 carbon atoms and the radicals R2, R3 and R4 are identical or different and are hydrogen or lower alkyl radicals, or in which the group NR2R3R4 is a cyclic ammonium group, and n has the value 0 or 1. Antitumor and antifungal activity are indicated for these compounds.
The present invention relates to alkyl or alkene phosphates in which the choline radical is part of a heterocyclic ring, to a process for the preparation of the class of compounds, to pharmaceutical compositions containing the compounds as active ingredients and to processes for the preparation of said drugs.
More specifically, the present invention provides compounds of the General Formula I: 
in which
R is a linear or branched alkyl radical having 10 to 24 carbon atoms, which can also contain one to three double or triple bonds, R1 and R2 independently of one another are hydrogen or in each case a linear, branched or cyclic saturated or unsaturated alkyl radical having 1 to 6 carbon atoms, which can also contain a Cl, OH or NH2 group, it also being possible for two of these radicals to be bonded together to form a ring,
A is a single bond or one of the groups of the formulae
xe2x80x94CH2xe2x80x94CH2xe2x80x94CH2xe2x80x94Oxe2x80x94xe2x80x83xe2x80x83(II),
xe2x80x94CH2xe2x80x94CH2xe2x80x94Oxe2x80x94xe2x80x83xe2x80x83(III), 
xe2x80x83the groups (II) to (VI) being orientated in such a way that the oxygen atom is bonded to the phosphorus atom of compound (I), X is an oxygen or sulphur atom or NH when A is a single bond, or an oxygen or sulphur atom when A is one of the groups (II) to (VI),
y is equal to 0 or a natural number between 1 and 3, and
m and n independently of one another are 0 or natural numbers, with the proviso that m+n=2 to 8.
The present invention also provides a pharmaceutical composition comprising, as the active ingredient, at least one compound according to General Formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier therefor. The pharmaceutical composition may also include pharmaceutically acceptable excipients, adjuncts, fillers and diluents. The amount of active ingredient in the pharmaceutical dosage unit the pharmaceutical composition is preferably between 50 mg and 250 mg. Preferred compounds for the pharmaceutical composition are selected from the group consisting of octadecyl 1,1-dimethylpiperidinio-4-yl phosphate, octadecyl 1,1-dimethylperhydroazepinio-4-yl phosphate, octaecyl 1,1-dimethylperhydroazepinio-4-yl phosphate, erucyl 1,1-dimethylpiperidinio-4-yl phosphate and erucyl 1,1-dimethylperhydroazepinio-4-yl phosphate.
The present invention also provides methods of treating a tumor, autoimmune disease or skin disease or skin disease, and of combating protozoal and fungal diseases, which comprises administering to a host in need o such treatment an effective amount of a compound of General Formula (I). Such methods are particularly useful for treating leishmaniasis, multiple sclerosis, and psoriasis.
In addition, the invention provides a method of treating bone marrow damage due to treatment with cytostatic agents and other myeloxtoxic active ingredients which comprises administering, to a host having bone marrow damage due to treatment with cytostatic agents or other myelotoxic active ingredients, an effective amount of a compound of General Formula (I).
The invention also provides a method of treating a viral disease which comprises administering to a host having such a disease an effective amount of a compound of General Formula (I). This method should be particularly useful in treating AIDS.
Surprisingly, the compounds according to the invention have better antitumor activity than the open-chain derivatives described in EP-A 108 565. The invention further relates to processes for the preparation and processes for the purification of the novel compounds.
More specifically, the present invention relates to a procedure for the preparation of compounds of general formula Ixe2x80x94further referred to as process Axe2x80x94in which a compound of the general formula VII.
Rxe2x80x94Xxe2x80x94Axe2x80x94Hxe2x80x83xe2x80x83(VII)
in which R, X and A are as defined above, is reacted with phosphorus oxytrichloride in the presence of a suitable auxiliary base, with or without a solvent, and then reacted with a compound of the general formula: 
in which R1, R2, y, m and n are as defined above and Yxe2x88x92 is halide, mesylate or tosylate, to give compounds of the general formula I, or optionally compounds of the General Formula IX: 
in which R1, y, m and n as defined above can be used instead of compounds of the general formula VIII during the process mentioned above. Process B consists in the subsequent alkylation of compounds of general formula I obtained by process A, in which R1 and/or R2 are hydrogen, using alkylating agents R2xe2x80x94Y in which R2 is as defined above and Y is chlorine, bromine, iodine, tosyl or mesyl, in a manner known per se.
The present invention also provides a process for the purification of the compounds of General Formula I in which a solution of the compounds of General Formula I, which have been prepared by means of known processes or by a process as described above, in an organic solvent is treated with a mixed bed ion exchanger or successively or simultaneously with an acid or basic ion exchanger.
The first step of process A consists in reacting phosphorus oxytrichloride with a compound of Formula VII in halogenated hydrocarbons, saturated cyclic ethers, acyclic ethers, saturated hydrocarbons having 5 to 10 carbon atoms or liquid aromatic hydrocarbons which can also be substituted by halogen (especially chlorine), or in mixtures of the above-mentioned solvents, or without a solvent, optionally in the presence of a basic substance conventionally used for this purpose.
Examples of possible halogenated hydrocarbons are hydrocarbons having 1 to 6 carbon atoms, one or more or all of the hydrogen atoms being replaced with chlorine atoms. Methylene chloride, chloroform, ethylene chloride, chlorobenzene and dichlorobenzene, for example, can be used. In the case of halogen-substituted aromatic hydrocarbons, these are preferably substituted by one or two halogen atoms.
Examples of saturated cyclic ethers which can be used are ethers with a ring size of 5-6 which consist of carbon atoms and one or 2 oxygen atoms, examples of such ethers being tetrahydrofuran and dioxane.
The acyclic ethers consist of 2 to 8 carbon atoms and are liquid, possible examples being diethyl ether, diisobutyl ether, methyl tert-butyl ether and diisopropyl ether.
Possible saturated hydrocarbons are unbranched and branched hydrocarbons which consist of 5 to 10 carbon atoms and are liquid, possible examples being pentane, hexane, heptane and cyclohexane.
Examples of possible aromatic hydrocarbons are benzene and alkyl-substituted benzenes, the alkyl substituents consisting of 1 to 5 carbon atoms.
Possible basic substances both for the reaction of the phosphorus oxychloride with the long-chain alcohol and for the subsequent conversion to the phosphoric acid diester are amines, for example aliphatic amines of the formula NR1R2R3, R1, R2 and R3 being identical or different and being hydrogen or C1-C6-alkyl, or else aromatic amines such as pyridine, picoline and quinoline. The basic substance required for the conversion to the phosphoric acid diester can be added simultaneously with or before the amino alcohol or ammonium alcohol salt.
A solvent is necessary in every case for the second reaction, i.e., if the first reaction step is carried out without a particular solvent, one must now be added. The molar ratio of phosphorus oxychloride to the long-chain alcohol is for example between 1.5:1 and 0.8:1.
The amino alcohol or the ammonium alcohol salt is for example used in excess, based on the long-chain alcohol (about 1.1-1.5 molar excess).
If the reaction of the phosphorus oxychloride with the long-chain alcohol is carried out in the presence of a basic substance, the amount of the basic substance is for example 1 to 3 mol, based on 1 mol of POCl3. The amount of basic substance used for the subsequent conversion to the phosphoric acid diester is for example 1 to 5 mol, based on 1 mol.
The temperature of the reaction of phosphorus oxychloride with the long-chain alcohol is between xe2x88x9230xc2x0 C. and +30xc2x0 C., preferably between xe2x88x9215xc2x0 C. and +5xc2x0 C. and especially between xe2x88x9210xc2x0 C. and xe2x88x925xc2x0 C.
The duration of this reaction is for example 0.5-5 hours, preferably 1-3 hours and especially 1.5-2 hours. If it is carried out in the presence of a basic substance, the reaction generally proceeds rapidly (about 30 minutes).
The amino alcohol or the ammonium alcohol salt is then added in portions or all at once. Possible ammonium alcohol salts are those with mineral acids (for example sulphuric acid, hydrochloric acid) and also those with organic acids, for example acetic acid, paratoluene-sulphonic acid and the like. This reaction step takes place in an inert solvent. Possible solvents for this step are the same ones as those used for the reaction of the phosphorus oxychloride with the long-chain alcohol, in the case where this reaction is carried out in a solvent..
The basic substance is then added dropwise, either dissolved in one of the indicated solvents or without a solvent. The following are preferably used as solvents for the basic substance: halogenated hydrocarbons, saturated cyclic ethers, acyclic ethers, saturated hydrocarbons having 5 to 10 carbon atoms, liquid aromatic hydrocarbons or mixtures of the above-mentioned solvents.
These are the same solvents as those which can be used for the reaction of the phosphorus oxychloride with the long-chain alcohol.
The addition of the basic substance raises the temperature. Care is taken to ensure that the temperature is kept in a range of between 0xc2x0 C. and 40xc2x0 C., preferably between 10xc2x0 C. and 30xc2x0 C. and especially between 15xc2x0 C. and 20xc2x0 C.
The reaction mixture is then stirred at 5xc2x0 C. to 30xc2x0 C., preferably 15xc2x0 C. to 25xc2x0 C. (for example for 1 hour to 40 hours, preferably 3 hours to 15 hours).
The reaction mixture is hydrolyzed by the addition of water, during which the temperature should be kept at between 10xc2x0 C. and 30xc2x0 C., preferably between 15xc2x0 C. and 30xc2x0 C. and especially between 15xc2x0 C. and 20xc2x0 C.
The above-mentioned hydrolyzing liquids can also contain basic substances, such basic substances possibly being alkali metal and alkaline earth metal carbonates and bicarbonates.
To complete the hydrolysis, stirring is then continued for a further 0.5 hour to 4 hours, preferably 1 to 3 hours and especially 1.5 to 2.5 hours, at 10xc2x0 C. to 30xc2x0 C., preferably at 15xc2x0 C. to 25xc2x0 C. and especially at 18xc2x0 C. to 22xc2x0 C.
The reaction solution is then washed with a mixture of water and alcohols (preferably saturated aliphatic alcohols having 1 to 4 carbon atoms) which can optionally also contain a basic substance. The mixing ratio water:alcohol can be for example between 5 and 0.5, preferably 1-3 (v/v).
Examples of possible basic substances for the washing liquid are alkali metal and alkaline earth metal carbonates and bicarbonates, as well as ammonia (for example aqueous ammonia). A 3% solution of sodium carbonate in water is particularly preferred.
The reaction solution can then optionally be washed with an acid solution. The acid washing is advantageous for removing basic components of the reaction solution which have not yet reacted, especially when methylene chloride is used as the solvent.
The washing solution consists of a mixture of water and alcohols. Mixtures of saturated aliphatic alcohols having 1 to 4 carbon atoms are preferred, it optionally being possible for an acid substance to be present as well. The mixing ratio water:alcohol can be for example between 5 and 0.5, preferably 1-3 (v/v).
Examples of possible acid substances for the washing liquid are mineral acids and organic acids, for example hydrochloric acid, sulphuric acid, tartaric acid or citric acid. A 10% solution of hydrochloric acid in water is particularly preferred.
This is followed by a further washing with a mixture of water and alcohols. Mixtures of saturated aliphatic alcohols having 1 to 4 carbon atoms are preferred, it optionally being possible for a basic substance to be present as well. The mixing ratio water:alcohol can be for example between 5 and 0.5, preferably 1-3.
The washed phases are then combined and dried in conventional manner, after which the solvent is removed (preferably under reduced pressure, for example at 5 to 100 mbar), optionally after the addition of 150-1000 ml, preferably 300-700 ml and especially 450-550 ml of an aliphatic alcohol (based on 1 molar part by weight of dry product). Preferred alcohols are saturated aliphatic alcohols with a chain length of 1 to 5 carbon atoms, particularly preferred alcohols being n-butanol and isopropanol. The purpose of this alcohol treatment is the complete removal of residual water and the avoidance of foaming.
Further purification of the product can be effected for example by dissolving the crude product in hot ethanol, filtering off the residue and treating the filtrate with a mixed bed ion exchanger such as, for example, Amberlite MB3 in ethanolic solution. Any commercially available acid and basic ion exchangers can be used, simultaneously or successively, instead of a mixed bed ion exchanger.
The solution is then recrystallized from ketones such as, for example, acetone or methyl ethyl ketone; digestion with the above solvents is sufficient in some cases. It may be convenient to purify the products by column chromatography or flash chromatography on silica gel using mixtures of chloroform, methylene chloride, methanol and 25% ammonia solution, for example, as the eluent.
Process variant B consists in the subsequent alkylation of products which are obtainable by process A using amino alcohols. Examples of alkylating agents which can be used are methyl p-toluenesulphonate or dimethyl sulphate. Possible solvents are those which have been mentioned above.
Alkali metal carbonates are examples of basic substances used. The reaction is carried out at elevated temperature, for example at the boiling point of the solvents.